Differential expression of a chloride intracellular channel gene, CLIC4, in transforming growth factor-β1-mediated conversion of fibroblasts to myofibroblasts
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Conversion of fibroblasts into myofibroblasts as mediated by transforming growth factor-β1 (TGF-β1) is the most prominent stromal reaction to a number of epithelial lesions including breast cancer. To identify genes which are regulated during this process, the mRNA profiles from primary breast fibroblasts treated with or without TGF-β1 were analyzed by differential display. Ninety-five differentially expressed transcripts were PCR-cloned and sequenced, and 28 clones were selected for verification in a hybridization array. By use of gene-specific sequence tags, nine differentially expressed genes were identified. One of the clones, identified as CLIC4, a member of the CLIC family of chloride channels, was up-regulated more than 16 times in myofibroblasts and was therefore chosen for further analysis. Using RT-PCR, comparison with CLIC1, CLIC2, CLIC3, and CLIC5 demonstrated that CLIC4 was unique by being up-regulated by TGF-β1 in myofibroblasts. Immunohistochemistry showed a hitherto unknown, distinctive pattern of CLIC4 expression in breast stroma. Whereas normal breast fibroblasts were devoid of CLIC4 protein expression, myofibroblasts of breast carcinomas were strongly CLIC4-positive. The functional significance of CLIC4 was analyzed in MEF/3T3 fibroblasts by conditional expression using the tetracycline-repressive gene regulation system. In a migration assay, we found that CLIC4 inhibited cell motility by 27%. These results suggest that CLIC4 is differentially regulated in fibroblasts and that its expression contributes to a collective stationary myofibroblast phenotype.
Originalsprog | Engelsk |
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Tidsskrift | American Journal of Pathology |
Vol/bind | 161 |
Udgave nummer | 2 |
Sider (fra-til) | 471-480 |
Antal sider | 10 |
ISSN | 0002-9440 |
DOI | |
Status | Udgivet - 2002 |
Bibliografisk note
Funding Information:
Supported by the Danish Research Council, the Danish Cancer Society, the Novo Nordic Foundation, the Meyer Foundation, the Friis Foundation, Fru Astrid Thaysens Legat for Lægevidenskabelig Grundforskning (to L.R.-J., R.V. and O.W.P.), Weimanns Legat (to L.R.-J.), National Institutes of Health grant CA-64786-02 (to O.W.P.), National Institutes of Health grant 44838 , and a Merit Award from the Department of Veterans Affairs (to J.C.E.).
ID: 304483750